中国激光, 2014, 41 (11): 1115002, 网络出版: 2014-10-08   

拉曼光谱技术研究铝胁迫下的土生隐球酵母细胞凋亡

Cell Apoptosis in Yeast under Aluminum Stress Analyzed by Laser Raman Spectroscopy
作者单位
1 昆明理工大学生物工程技术研究中心, 云南 昆明 650500
2 广西科学院生物物理实验室, 广西 南宁 530007
摘要
运用激光镊子拉曼光谱技术(LTRS)研究铝胁迫下土生隐球酵母细胞和线粒体拉曼光谱变化。细胞和线粒体中与核酸、蛋白质和脂类相关的特征峰强度随着铝处理浓度和处理时间的增加而逐渐降低,表明细胞凋亡过程中细胞和线粒体内的核酸、蛋白质和脂类生物大分子逐渐减少。线粒体内与细胞色素c(Cyt c)相关的特征峰强度随铝处理浓度增大和处理时间的延长而显著降低,说明线粒体中的细胞色素c释放到线粒体外。线粒体的呼吸峰随着铝浓度的增加和处理时间的延长而降低,说明线粒体的活性不断减弱,能量代谢受阻。因此,拉曼光谱实时监测了铝胁迫细胞凋亡的动态过程及其线粒体的生理生化变化过程,并且揭示了铝诱导土生隐球酵母凋亡过程中线粒体内细胞色素c的释放行为,这些结果有助于了解酸性土壤中铝对生物的毒害机理。
Abstract
Laser Raman spectroscopy (LTRS) is used to monitor the Raman spectroscopic changes of Cryptococcus humicolus cells and mitochondria stressed by aluminum (Al). The results show that the characteristic Raman peaks of C. humicolus cells and mitochondria are significantly decreased with the increase of aluminum concentration and treatment time, the intensity of Raman peaks associated to nucleic acid, protein and lipid respectively exhibits significant decreasing tendency with the extension of treatment time, indicating that the content of nucleic acid, protein and lipid in the cells and mitochondria is reduced during the apoptosis process. The intensity of characteristic peaks related to cytochrome c (Cyt c) in mitochondria significantly decreases with the increase of aluminum concentration and treatment time, suggesting that the yeast mitochondrial membrane rupture leads to Cyt c release into outer of mitochondria. With the increase of aluminum concentration and treatment time, the respiratory peak is observed to decrease gradually, indicating that the mitochondrial activity becomes weaker, energy metabolism has been hampered. LTRS is used for real-time monitoring of the dynamic process of cell apoptosis and physiological and biochemical changes of in vitro mitochondria under aluminum stress, and revealing the release behavior of Cyt c during the apoptosis process of C. humicolus cells induced by aluminum. The results help understand the biological mechanism of aluminum toxicity in acidic soils on organisms.

李金金, 卢明倩, 张晶晶, 黄庶识, 陈丽梅, 年洪娟. 拉曼光谱技术研究铝胁迫下的土生隐球酵母细胞凋亡[J]. 中国激光, 2014, 41(11): 1115002. Li Jinjin, Lu Mingqian, Zhang Jingjing, Huang Shushi, Chen Limei, Nian Hongjuan. Cell Apoptosis in Yeast under Aluminum Stress Analyzed by Laser Raman Spectroscopy[J]. Chinese Journal of Lasers, 2014, 41(11): 1115002.

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